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Determination of Aromatic and Branched-Chain Amino Acids in Plasma by HPLC With Electrogenerated Ru(bpy)3(3+) Chemiluminescence Detection

Determination of Aromatic and Branched-Chain Amino Acids in Plasma by HPLC With Electrogenerated Ru(bpy)3(3+) Chemiluminescence Detection

Kazuo Uchikura

Chem Pharm Bull (Tokyo). 2003 Sep;51(9):1092-4.

PMID: 12951455

Abstract:

An HPLC method is described for the electrochemiluminescence (ECL) detection of amino acids, following cycloaddition reaction of their amino groups with divinyl sulfone (DVS), using electrogenerated tris(bipyridine)ruthenium(III). The derivatization reaction conditions were examined, with the optimum conditions found to be 40 mM DVS (pH 8.0) at 50 degrees C for 15 min. Detection limits for the 15 amino acids examined varied greatly (0.04-8.0 pmol) using a standard solution by flow injection analysis (FIA). These optimized conditions were used for HPLC determination of the amino acids in human plasma. A linear relationship was obtained up to 100 pmol on a column for aromatic and branched-chain amino acids. Recoveries of Tyr, Met, Val, Leu, Ile, Phe and Trp when added to human plasma (1 micromol/10 ml plasma, n=5) were 101.5+/-1.1, 99.0+/-1.2, 98.0+/-1.4, 101.1+/-1.6, 95.1+/-1.6, 99.2+/-1.5 and 97.7+/-1.3 % (mean+/-S.D.) respectively. The concentrations of the amino acids in the plasma are in good agreement with other published data.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AS212795	FIA Aromatics Standard			Price

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